The objective of Project 2 is to provide non-clinical safety data and pharmacological evidence of a Chinese[unreadable] herbal preparation used in the treatment of irritable bowel syndrome (IBS). To achieve this objective, this[unreadable] Project proposes to carry out toxicity tests and pharmacological studies of the prescription. Toward this end,[unreadable] in vitro and in vivo models will be established for toxicity and pharmacological evaluations. Toxicity tests[unreadable] include acute (single-dose), subacute (multiple-dose for 14 days), and chronic toxicity (multiple-dose for 3[unreadable] months) tests in rodents, mutagenicity tests using bacteria (Ames test), animal cells (chromosome aberration[unreadable] test), as well as intact animals (micronucleus test). Functional assays for in vitro intestinal contractility will be[unreadable] set up to test for anticholinergic (against acetylcholine), antiserotonic (against histamine), antimuscarinic (M3[unreadable] antagonism), and opioid agonistic effects of the herbal preparation as well as individual plant extracts.[unreadable] Receptor binding assays will be used to confirm the effects if necessary. The therapeutic potential of the[unreadable] Chinese medicinal recipe will be evaluated in an animal model of visceral hyperalgesia induced by neonatal[unreadable] maternal separation. In addition we will provide bioassay facilities to aid the isolation of active principles from[unreadable] the medicinal herbs. At the end of this study, it is anticipated that (1) Non-clinical safety data will be generated[unreadable] to supplement human traditional experience in defining safe use of the prescription, and thus supporting the[unreadable] clinical applications; (2) Pharmacological actions of the prescription will be characterized so as to provide a[unreadable] better insight for further development of the herbal prescription.[unreadable]